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. 2023 Jun 15;18(7):1451–1467. doi: 10.1016/j.stemcr.2023.05.012

Figure 3.

Figure 3

Enteral Ala-Gln supplementation augments the activity of ex vivo ISCs to form enteroids

(A) Representative images of the morphology of primary enteroids expanded from crypt cells at 24, 48, 72, 96, and 120 h are shown. Scale bars, 200 μm.

(B) Enteroid-forming capacity of mice jejunum crypts (n = 16, 16 wells of separated crypts from 4 independent mice per condition were analyzed, means ± SD; ∗p < 0.05, ∗∗p < 0.01; t test, results of 3 independent experiments).

(C and D) Quantification of crypt domains (C) and surface area (D) per primary enteroids at 96 h (n = 24, 24 enteroids from 4 independent mice per condition were analyzed, means ± SD; ∗p < 0.05, ∗∗p < 0.01; t test, results of 3 independent experiments).

(E) Representative images of the morphology of secondary enteroids expanded from crypt cells at 12, 36, 60, and 84 h are shown. Scale bars, 200 μm.

(F and G) Quantification of crypt domains (F) and surface area (G) per secondary enteroid (n = 18, 18 enteroids from 3 independent mice per condition were analyzed, means ± SD; ∗p < 0.05, ∗∗p < 0.01; t test, results of 3 independent experiments).

(H) Representative images of primary enteroids of immunofluorescence stained with OLFM4 (green) and DAPI (blue) are shown. Scale bars, 100 μm.

(I) Western blot analysis of intestinal stem cells marker OLFM4 protein expression in primary enteroids (n = 3, primary enteroids from 3 independent mice per condition, results of 3 independent experiments).